Rupture disks for bioreactors and methods of using same

ABSTRACT

A rupture disk includes a body including an internal wall extending from a first end of the body to a second end of the body. The internal wall defines an opening extending through the body. In some embodiments, the body further includes an internal flange formed on the internal wall, and the internal flange has a stepped surface. A rupture membrane is positioned within the body, and secured, for example, by cold welding. The rupture membrane may be secured to the stepped surface. The rupture membrane is positioned within the body such that the rupture membrane does not extend beyond the first end of the body. A process system including the rupture disk is also provided. A method of maintaining a process system that includes the rupture disk is also provided.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to U.S. ProvisionalPatent Application Ser. No. 62/431,985 filed on Dec. 9, 2016, which ishereby incorporated herein by reference in its entirety for allpurposes.

FIELD OF THE INVENTION

The disclosure relates to rupture disks and methods of using and makingthe same.

BACKGROUND

The manufacture of biological products includes many processes that areconducted in closed systems or pressurized systems. In addition tobioreactors, conduits, valves, reservoirs and the like such systemstypically include components that provide protection from deviationsfrom operating pressure, e.g., over-pressurization. Rupture discs arenon-reclosing pressure relief devices. They can provide relieve ifsystem pressure, e.g., vessel or bioreactor pressure, is to low or toohigh. Rupture discs typically provide instant response (withinmilliseconds) to an increase or decrease in system pressure. Once therupture disc has ruptured it will not reseal, it is “sacrificial.”Typically a rupture disk includes a one-time-use membrane that fails ata predetermined differential pressure, either positive or vacuum. Atypical rupture disk is made of metal and is configured as a thin,circular membrane that is held firmly in a holder or housing, with anelastomeric member providing a hermetic seal between the rupture diskand housing.

Rupture occurs when the fluid pressure within the vessel or systemexceeds the design rupture pressure of the disk. This causes fluidpressure to be relieved from the vessel or system.

In bioprocess systems, rupture disks are typically installed on a spoolpiece between a tank and a vent tube. Typically elastomeric members areused to seal the union of the rupture membrane and its housing.

Rupture disks are commonly used in series (upstream) with a relief valveto prevent corrosive fluids from contacting the metal parts of thevalve.

SUMMARY

Sealing members, which are typically made of elastomeric materials, needperiodic or event-based replacement. In particular, the need to replace,e.g., due to failure or scheduled maintenance, the elastomeric memberthat joins the rupture membrane to the housing of a rupture diskassembly can result in the need to replace the entire rupture diskassembly, even though the rupture disk is otherwise still serviceable.Even if the elastomeric member that joins the rupture disk to thehousing could be replaced, such replacement would involve substantialmanipulation of the rupture disk and could lead to premature failure ofthe rupture disk. Thus, replacement of the sealing member that joins therupture disk to the housing of the rupture disk assembly typicallyrequires replacement of the entire rupture disk assembly, including theotherwise serviceable rupture membrane and housing. It has beendiscovered that the sealing member that joins the rupture membrane tothe housing of the rupture disk can be eliminated by providing a directintegral hermetic joint, e.g., such as is formed by cold welding,between the rupture membrane and the housing of a rupture disk assembly.The need to replace rupture disk assemblies is minimized because thereis no sealing member to replace. The rupture disk assembly can stillrely on sealing members to provide a seal between the rupture diskassembly and other components, e.g., tanks, but upon replacement of suchsealing members the rupture disk assembly can be reused.

According to an aspect of the present disclosure, a rupture diskcomprises a body including an internal wall extending from a first endof the body to a second end of the body, the internal wall defining anopening extending through the body, the body further including aninternal flange formed on the internal wall, the internal flange havinga stepped surface; and a rupture membrane positioned within the body,the rupture membrane including a rupture membrane surface secured to thestepped surface to form a hermetic seal, the stepped surface and therupture membrane being positioned within the body such that the rupturemembrane does not extend beyond the first end of the body.

In some embodiments, the body is configured for removable attachment toa component.

In some embodiments, the body includes a first metal and the rupturemembrane is fabricated from a second metal, wherein the first metal isselected from 316L stainless steel, Inconel, indium, aluminum, copper,lead, zinc, nickel, silver, palladium, cadmium, titanium, tungsten, tin,gold, and lead, and wherein the second metal is selected from 316Lstainless steel, Inconel, indium, aluminum, copper, lead, zinc, nickel,silver, palladium, cadmium, titanium, tungsten, tin, gold, and lead.

In some embodiments, the stepped surface of the internal flange and therupture membrane surface of the rupture membrane are joined together bya metal-to-metal bond.

In some embodiments, the stepped surface of the internal flange and therupture membrane surface of the rupture membrane are joined together bya cold weld.

In some embodiments, the rupture disk is configured to form a barrierthat seals an aperture of a component to which the rupture disk isattached, and wherein the rupture membrane, prior to rupture, preventsflow of a fluid out of the aperture of the component.

In some embodiments, the aperture is greater than 10 mm.

In some embodiments, the component is suitable for use in the productionof a biological product.

In some embodiments, the body is configured for attachment to acomponent by a connector.

In some embodiments, the component comprises a chamber or reservoir orother void, the chamber or reservoir or other void having a lineardimension of greater than 20 centimeters.

In some embodiments, the rupture disk is configured to be disposedbetween a first and second component, and wherein the rupture membrane,prior to rupture, forms a barrier between the first component and thesecond component.

In some embodiments, the body is configured as a spool piece.

In some embodiments, the body is configured so as to be removablyattached by a clamp to a component.

In some embodiments, the rupture disk further comprises a gasketconfigured to be disposed between the first end of the body and acomponent and to provide a fluid-proof seal there between.

In some embodiments, the rupture membrane is positioned such that it isentirely within the body.

In some embodiments, the rupture membrane is positioned such that it isentirely within the body, but close enough to the first end of thehousing to minimize a volume defined by a clean surface of the rupturemembrane and a portion of the internal wall between the clean surface ofthe rupture membrane and the first end of the body.

In some embodiments, the rupture membrane is positioned such that it isentirely within the body such that a volume of a space defined by theface of the rupture membrane and a portion of the internal wall betweenthe face of the rupture membrane and the first end of the body is nomore than 10 cm³.

In some embodiments, the internal flange and the internal wall areintegrally formed.

In some embodiments, the rupture disk consists of the rupture membraneand the body formed as a spool piece.

In some embodiments, the rupture disk consists essentially of therupture membrane and the body formed as a spool piece.

According to another aspect of the present disclosure, a process systemcomprises a rupture disk of the present disclosure, wherein the processsystem includes a bioreactor. In some embodiments, the process system isa bioprocess system.

According to another aspect of the present disclosure, a method of amethod of maintaining a process system comprises detaching a rupturedisk of claim 1, from a component; and reattaching the rupture disk tothe component, thereby maintaining the process system. In someembodiments, the process system is a bioprocess system.

In one aspect, the disclosure features a rupture disk assembly includinga housing; and a rupture membrane, wherein the rupture membrane isdisposed within the housing, e.g., is integral with the housing, and thejoint between the housing and rupture membrane is hermetic.

In an embodiment, the housing is configured for removable attachment toa second component, e.g., attachment by a mechanical fastener, e.g., apressure or torque-based fastener, e.g., a bolt or clamp.

In an embodiment, the rupture disk assembly is configured for attachmentto a second component with a hygienic clamp.

In an embodiment, the housing is configured for removable attachment toa second component, e.g., attachment by a mechanical fastener, e.g., apressure or torque based fastener, e.g., a bolt or clamp.

In an embodiment, the housing is configured for attachment to a secondcomponent, by other than a weld, e.g., a cold weld.

In an embodiment, the housing is further configured for removableattachment to a third component, e.g., attachment by a mechanicalfastener, e.g., a pressure or torque-based fastener, e.g., a bolt orclamp.

In an embodiment, the rupture disk assembly is further configured forattachment to a third component with a hygienic clamp.

In an embodiment, the housing is further configured for removableattachment to a third component, e.g., attachment by a mechanicalfastener, e.g., a pressure or torque based fastener, e.g., a bolt orclamp.

In an embodiment, the housing is further configured for attachment to athird component, by other than a weld, e.g., a cold weld. In anembodiment, the rupture disk assembly includes a cold-welded jointbetween the housing and the rupture membrane. In an embodiment therupture membrane is positioned such that it is entirely within thehousing, and e.g., if concave does not protrude beyond the face of thehousing.

In an embodiment the rupture membrane is positioned such that it isentirely within the housing but close enough to the face of the housingto minimize volume of the space defined by the face of the rupturemembrane and the face of the housing that is adjacent a first component.

In an embodiment the rupture membrane is positioned such that it isentirely within the housing but close enough to the face of the housingsuch that volume of the space defined by the face of the rupturemembrane and the face of the housing that is adjacent a first componentis no more than 5, 10, 15, 20, 30, 40, 50, 100, 200, 300, 400, or 500cm³.

In an embodiment the rupture membrane has a diameter, or longestdimension of, 1-100 cm, 1-50 cm, 1-25 cm, 1-10 cm to 5 cm.

In an embodiment the rupture membrane has a diameter, or longestdimension of, 5-100 cm, 5-50 cm, 5-25 cm, or 5-10 cm.

In an embodiment, the housing in the rupture disk assembly comprises ametal component and the rupture membrane is metal.

In an embodiment, the metal component and the rupture membrane comprisethe same metal.

In an embodiment, the metal component comprises a first metal and therupture membrane comprises a second metal. In an embodiment, the firstmetal, the second metal, or both, metals comprise stainless steel (e.g.,SAE 304 or SAE 316L stainless steel), carbon steel, gold, platinum, or acombination thereof.

In an embodiment, the metal component and rupture membrane are joined bycold-welding. In an embodiment, a surface of the rupture membrane and asurface of the housing are joined together by a metal-to-metal bond.

In an embodiment, the metal-to-metal bond is formed without substantialheat input, e.g., in the absence of heat that results in a molten orliquid phase at the joint in the joining, e.g., a welding process.

In an embodiment, the housing comprises a component including, e.g., amaterial selected from silicon, metals, ceramics, polymers, glasses, andcombinations thereof.

In an embodiment, the rupture disk assembly is configured to form abarrier that seals an aperture of a second component to which therupture disk assembly is attached and prior to rupture prevents flow ofa fluid, e.g., a liquid or gas, or mixture of a liquid and a gas, out ofthe second component.

In an embodiment, the second component comprises a vessel, e.g., apressure vessel.

In an embodiment, the second component comprises a vessel required bylaw, statute, rule or regulation, to have a pressure relief component.

In an embodiment, the rupture disk assembly includes an aperture greaterthan 2, 3, 4, 5, 10, 15, or 20 mm.

In an embodiment, the second component is a component used in theproduction of a biological product, e.g., a polypeptide, e.g., acomponent or biological product described herein.

In an embodiment, the second component comprises a chamber or reservoirin liquid communication with the rupture membrane, e.g., the chamber orreservoir vessel.

In an embodiment, the chamber or reservoir vessel has a capacity orvolume of at least 0.1, 0.5, 1.0, 10, 20, 30, 40, 50, 100, 200, 500,1,000, 1,500, 10,000, 12,500, 15,00, 17,500, 20,000, 25,000 or 30,000 L.

In an embodiment, the second component comprises a wall or housing,e.g., defining at least a portion of the chamber or reservoir. In anembodiment, the thickness of the wall or housing is greater than 1, 10,5, 10, 15, or 20 mm.

In an embodiment, the housing of the rupture disk assembly is configuredfor attachment to the second component by a joint that is not integral,e.g., a joint that including a sealing member, e.g., a deformablemember, e.g., a gasket, e.g., an elastomeric gasket.

In an embodiment, the second component comprises a chamber or reservoiror other void, the chamber or reservoir or other void having a lineardimension, e.g., depth, diameter or radius, of greater than 3, 4, 5, 10,20, 30, 50, 100 centimeters.

In an embodiment, the rupture disk assembly is configured so as to bedisposed between a first and second component and wherein the rupturemembrane, prior to rupture, forms a barrier between the second componentand a third component.

In an embodiment, the rupture disk assembly is configured so as to bedisposed between a second and third component. In an embodiment, therupture membrane, prior to rupture, prevents flow of fluid, e.g., aliquid or gas, or mixture of a liquid and a gas, from the secondcomponent into the third component.

In an embodiment, the second or third component of the rupture diskassembly, e.g., a vessel, has a capacity or volume of at least 0.1, 0.5,1.0, 10, 20, 30, 40, 50, 100, 200, or 500 liters of liquid.

In an embodiment, the housing is configured as a spool piece.

In an embodiment, the housing of the rupture disk assembly is configuredas a cylinder having a lumen and the rupture membrane is disposed so asto separate a first portion of the lumen from a second portion of thelumen.

In an embodiment, the housing of the rupture disk assembly is configuredso as to be attached to a second component, e.g., a vessel, e.g., atank, or a pipe.

In an embodiment, the housing of the rupture disk assembly is not weldedto a second component, e.g., a vessel, e.g., a tank, or a pipe.

In an embodiment, the housing of the rupture disk assembly is configuredso as to be removably attached, e.g., attached by a bolt, clamp, orother removable fastener, to a second component, e.g., a vessel, e.g., atank, or a pipe.

In an embodiment, the housing of the rupture disk assembly is configuredfor disposition or placement of a sealing member, e.g., a deformablemember, e.g., a gasket, e.g., an elastomeric gasket, between the housingand a second component.

In an embodiment, the housing of the rupture disk assembly is configuredfor disposition or placement of a sealing member, e.g., a deformablemember, e.g., a gasket, e.g., an elastomeric gasket, between the housingand a third component.

In an embodiment, the housing of the rupture disk assembly is configuredto be attached, e.g., by a fastener, e.g., by a clamp or bolt, to asecond component, e.g., a conduit or pipe, pipe spool, flange, a valve,a reversible pressure regulator, a heat exchanger, or a vessel, e.g., abioreactor.

In an embodiment, the rupture disk assembly further comprises a sealingmember, e.g., a deformable member, e.g., a gasket, e.g., an elastomericgasket, configured to be disposed between the housing and the secondcomponent and provide a fluid-proof seal there between.

In an embodiment, the housing of the rupture disk assembly is integralwith e.g., welded to, a second component, e.g., a conduit or pipe, pipespool, flange, a valve, a reversible presser regulator, or a vessel,e.g., a bioreactor, provided that the second component has a capacity orvolume of at least 0.1, 0.5, 1.0, 10, 20, 30, 40, 50, 100, 200, or 500liters of liquid or is attached directly or indirectly to anothercomponent having such volume or capacity.

In an embodiment, the rupture disk assembly further comprises apackaging component configured so as to protect the rupture membranefrom damage or unwanted contact with other objects. Typically, thepackaging component protects the pressure-side face of the rupturemembrane. For example, the rupture disk assembly can comprise a cap,e.g., a polymeric or plastic cap, configured to fit over the aperture ofthe housing nearest the rupture membrane. The cap can be configured soas to fit tightly but removably from the rupture disk assembly. Therupture disk assembly can comprise a second cap configured and placed soas to protect the non-pressure-side face of the rupture membrane.

In an embodiment, the rupture disk assembly of the process system isdisposed between a first component and a second component.

Also provided is a process system including a rupture disk assembly asdescribed above, and a second component, e.g., a second componentdescribed herein, e.g., wherein the rupture disk assembly and the secondcomponent are in fluid connection.

In an embodiment of the process system, the second component comprises avessel, e.g., a bioreactor, a conduit, e.g., a pipe, a valve, e.g., areversible pressure regulator, an element for providing a readout ofpressure, e.g., upstream or downstream of the rupture disk assembly.

In an embodiment of the process system, the second component, e.g., avessel, has a capacity or volume of at least 0.1, 0.5, 1.0, 10, 20, 30,40, 50, 100, 200, or 500 liters, of liquid.

In an embodiment of the process system, the rupture disk assembly isattached to a second component.

In an embodiment of the process system, the rupture disk assembly isattached to a second component by a mechanical fastener, e.g., by clampsor bolts.

In an embodiment, the process system further includes a sealing member,e.g., a deformable member, e.g., a gasket, e.g., an elastomeric gasket,disposed between the housing and the second component.

In an embodiment, the process system includes a rupture disk assemblywith a housing that is integral with. e.g., welded to, a secondcomponent, e.g., a bioreactor, a conduit, e.g., a pipe, a valve, e.g., areversible pressure regulator, an element for providing a readout ofpressure, e.g., upstream or downstream of the rupture disk assembly,provided that the second component has a capacity or volume of at least0.1, 0.5, 1.0, 10, 20, 30, 40, 50, 100, 200, or 500 liters of liquid oris attached directly or indirectly to another component having suchvolume or capacity.

In an embodiment, the process system includes a second component thatcomprises a vessel, e.g., a bioreactor. In an embodiment, the secondcomponent comprises a valve, e.g., a reversible pressure regulator.

In an embodiment, one or both of the second and third component isdirectly attached to the rupture disk assembly.

In an embodiment, a fourth component is disposed between the rupturedisk assembly and the second or third component.

Also provided is a method of making a rupture disk assembly thatincludes forming a direct (e.g., lacking a third component such as anelastomer) hermetic joint between a rupture membrane and a housing,thereby making a rupture disk assembly.

In an embodiment, the method includes forming a cold-weld joint betweenthe rupture disk membrane and the housing.

In an embodiment, the rupture disk assembly is a rupture disk assemblyas herein described.

In an embodiment, the housing is configured for removable attachment toa second component, e.g., attachment by a mechanical fastener, e.g., apressure or torque-based fastener, e.g., a bolt or clamp.

In an embodiment, the rupture disk assembly is configured for attachmentto a second component with a hygienic clamp.

Also provided by the disclosure is a method of maintaining a processsystem, e.g., a process system as herein described, comprising detachinga rupture disk assembly, e.g., a rupture disk assembly as hereindescribed, from a second component, e.g., a vessel, e.g., a reactorvessel, and, optionally, inspecting, maintaining, or replacing a sealingmember disposed between the rupture disk assembly and the secondcomponent. The method further includes reattaching the rupture diskassembly to a second component, e.g., the second component from which itwas detached, thereby maintaining a process system.

Also provided the disclosure is a method of making a rupture diskassembly that includes forming a direct (e.g., lacking a third componentsuch as an elastomer) hermetic joint between a rupture membrane and ahousing, thereby making a rupture disk assembly.

In an embodiment, the method includes forming a cold-weld joint betweenthe rupture disk membrane and the housing.

In an embodiment, the rupture disk assembly is a rupture disk assemblyas herein described.

In another aspect, the disclosure provides a method of replacing agasket or sealing member-based rupture disk, comprising identifying arupture disk or rupture disk assembly to be replaced; removing theto-be-replaced rupture disk or rupture disk assembly, and installing anew rupture disk assembly.

Also provided by the disclosure is a method of maintaining a processsystem, e.g., a process system as herein described, comprising detachinga rupture disk assembly, e.g., a rupture disk assembly as hereindescribed, from a second component, e.g., a vessel, e.g., a reactorvessel, and, optionally, inspecting, maintaining, or replacing a sealingmember disposed between the rupture disk assembly and the secondcomponent. The method further includes reattaching the rupture diskassembly to a second component, e.g., the second component from which itwas detached, thereby maintaining a process system.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a cross-sectional view of an embodiment of a rupture diskassembly;

FIG. 2 is a perspective view of an embodiment of the rupture diskassembly;

FIG. 3A shows a rupture disk housing;

FIG. 3B shows a rupture membrane carrier; and

FIG. 3C shows the rupture membrane carrier secured within the rupturedisk housing to form an assembled rupture disk assembly.

DETAILED DESCRIPTION

The invention provides rupture disk assemblies that provide a sealbetween the rupture disk or membrane and a housing that is integral anddoes not include a separated sealing member, and methods of making andusing the same. In embodiments the rupture disk assembly is not integralwith the vessel, e.g., tank, it protects.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the invention pertains. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice of and/or for the testing of the present invention, thepreferred materials and methods are described herein. In describing andclaiming the present invention, the following terminology will be usedaccording to how it is defined, where a definition is provided.

It is also to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto be limiting. The articles “a” and “an” are used herein to refer toone or to more than one (i.e., to at least one) of the grammaticalobject of the article. By way of example, “a cell” can mean one cell ormore than one cell.

As used herein, “cold welding” refers to a process of joining a firstand a second metal surface without fusion/heating and without theproduction of a liquid or molten phase at the site of the weld.Typically, mating the two surfaces at high pressure plays a key role incold welding.

As used herein, the term “hermetic seal or joint” refers to seal thatprevents a fluid, e.g., a liquid or gas, from passing.

As used herein a “rupture disk or membrane” refers to a device which,prior to reaching the failure pressure, obstructs the path of a liquid.At failure pressure, the device opens (by irreversible cleavage orseparation-based failure) to allow flow of the liquid. In an embodimentthe rupture disk ruptures or fails at a predetermined differentialpressure, either positive or vacuum.

As used herein, the terms “peptide,” “polypeptide,” and “protein” areused interchangeably, and refer to a compound comprised of amino acidresidues covalently linked by peptide bonds, or by means other thanpeptide bonds. A protein or peptide must contain at least two aminoacids, and no limitation is placed on the maximum number of amino acidsthat can comprise a protein's or peptide's sequence. In one embodiment,a protein may comprise of more than one, e.g., two, three, four, five,or more, polypeptides, in which each polypeptide is associated toanother by either covalent or non-covalent bonds/interactions.Polypeptides include any peptide or protein comprising two or more aminoacids joined to each other by peptide bonds or by means other thanpeptide bonds. As used herein, the term refers to both short chains,which also commonly are referred to in the art as peptides,oligopeptides and oligomers, for example, and to longer chains, whichgenerally are referred to in the art as proteins, of which there aremany types. “Polypeptides” include, for example, biologically activefragments, substantially homologous polypeptides, oligopeptides,homodimers, heterodimers, variants of polypeptides, modifiedpolypeptides, derivatives, analogs, fusion proteins, among others.

“Product” as that term is used herein refers to a molecule, nucleicacid, polypeptide, or any hybrid thereof, that is produced, e.g.,expressed, by a cell which has been modified or engineered to producethe product. In one embodiment, the product is a naturally occurringproduct or a non-naturally occurring product, e.g., a synthetic product.In one embodiment, a portion of the product is naturally occurring,while another portion of the product is non-naturally occurring. In oneembodiment, the product is a polypeptide, e.g., a recombinantpolypeptide. In one embodiment, the product is suitable for diagnosticor pre-clinical use. In another embodiment, the product is suitable fortherapeutic use, e.g., for treatment of a disease. In one embodiment,the modified or engineered cells comprise an exogenous nucleic acid thatcontrols expression or encodes the product. In other embodiments, themodified or engineered cells comprise other molecules, e.g., that arenot nucleic acids, that controls the expression or construction of theproduct in the cell.

“Recombinant polypeptide” or “recombinant protein” as those terms areused herein refer to a polypeptide that can be produced by a celldescribed herein. A recombinant polypeptide is one for which at leastone nucleotide of the sequence encoding the polypeptide, or at least onenucleotide of a sequence which controls the expression of thepolypeptide, was formed by genetic engineering (of the cell or of aprecursor cell). E.g., at least one nucleotide was altered, e.g., it wasintroduced into the cell or it is the product of a geneticallyengineered rearrangement. In an embodiment, the sequence of arecombinant polypeptide does not differ from a naturally occurringisoform of the polypeptide or protein. In an embodiment, the amino acidsequence of the recombinant polypeptide differs from the sequence of anaturally occurring isoform of the polypeptide or protein. In anembodiment, the recombinant polypeptide and the cell are from the samespecies. In an embodiment, the recombinant polypeptide is endogenous tothe cell, in other words, the cell is from a first species and therecombinant polypeptide is native to that first species. In anembodiment, the amino acid sequence of the recombinant polypeptide isthe same as or is substantially the same as, or differs by no more than1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% from, a polypeptideencoded by the endogenous genome of the cell. In an embodiment, therecombinant polypeptide and the cell are from different species, e.g.,the recombinant polypeptide is a human polypeptide and the cell is anon-human, e.g., a rodent, e.g., a CHO, or an insect cell. In anembodiment, the recombinant polypeptide is exogenous to the cell, inother words, the cell is from a first species and the recombinantpolypeptide is from a second species. In one embodiment, the polypeptideis a synthetic polypeptide. In one embodiment, the polypeptide isderived from a non-naturally occurring source. In an embodiment, therecombinant polypeptide is a human polypeptide or protein which does notdiffer in amino acid sequence from a naturally occurring isoform of thehuman polypeptide or protein. In an embodiment, the recombinantpolypeptide differs from a naturally occurring isoform of the humanpolypeptide or protein at no more than 1, 2, 3, 4, 5, 10, 15 or 20 aminoacid residues. In an embodiment, the recombinant polypeptide differsfrom a naturally occurring isoform of the human polypeptide by no morethan 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15% of its amino acid residues.As used herein a “hygenic clamp” or “sanitary clamp”, refers to a clampconfigured to attach a rupture disk assembly to another component,wherein the clamp prevents exposure of the contents of the fluid (liquidor gas) flowing through the rupture disk assembly to the exterior.Exemplary hygienic clamps include sanitary, leak-proof fittings known inthe art, including the TRI-CLOVER® fitting sold by Alfa Laval, Inc.(Richmond, Va. USA). In embodiments, the hygienic clamp has dimensionsdisclosed in ASME Table DT-5-2 (“Hygenic Clamp Ferrule StandardDimensions and Tolerances”, 2009), which is incorporated herein byreference in its entirety.

Rupture Disk Assemblies

One embodiment of a rupture disk assembly (or rupture disk) 10 isdepicted and generally indicated in the cross-section of FIG. 1 and inthe perspective view in FIG. 2. As shown, the rupture disk assembly 10includes a housing 12 that is configured with a body portion 14, a firstflange 16 that has a first face 18, and a second flange 20 that has asecond face 22. Each of the first face 18 and the second face 22 issuitable for attaching the rupture disk 10 to other components. Thefirst face 18 can include one or more openings 24 and the second face 22can include one or more openings 26 for attaching the rupture diskassembly 10 to other components, including other components not shown inFIGS. 1 and 2. For example, the body portion 14 is configured forremovable attachment to a component, such as a bioprocess container. Insome embodiments, the body portion 14 is configured to be attached to acomponent by a connector, such as a clamp, a sanitary clamp, a bolt, apressure-based fastener, a torque-based fastener, a hygienic clamp, amechanical fastener, or another fastener.

The first face 18 and the second face 22 are each suitable for matingwith an elastomeric sealing member, such as an elastomeric o-ring, toprovide a seal between the rupture disk assembly 10 and one or morecomponents to which the rupture disk assembly 10 is attached. In someembodiments, the body portion 14 is configured to be removably attachedby a clamp to a component. In some embodiments, a sealing member, isdisposed between a first end 30 of the body portion 14 and a componentto which the body portion 14 is attached to provide a fluid-proof sealbetween the body portion 14 and the component to which the body portion14 is attached. In some embodiments, the sealing member may be a gasket31 configured to be disposed between the first end 30 of the bodyportion 14 and a component to which the rupture disk 10 is secured, andto provide a fluid-proof seal there between.

The body portion 14 includes an internal wall 32 that extends from thefirst end 30 of the body portion 14 to a second end 34 of the bodyportion 14. The internal wall 32 defines an opening that extends throughthe body portion 14. In some embodiments, the internal wall 32 defines astraight bore opening that extends through the body portion 14.

The internal wall 32 is configured to support a rupture membrane 40. InFIG. 1, the body portion 14 includes an internal flange 42 that isformed on the internal wall 32. The internal flange 42 may be formed,for example, by machining the internal wall 32 and the internal flange42 when forming the body portion 14. In other embodiments, the internalflange 42 may be secured to the internal 32 wall of the body portion 14,for example by cold welding, to form a hermetic seal between theinternal flange 42 and the body portion 14. The internal flange 42 has astepped surface 44 that is configured to support the rupture membrane40, as described further herein.

In some embodiments, the internal flange 42 and the internal wall 32 areintegrally formed. In some embodiments, the internal flange 42 and theinternal wall 32 are an integral structure. In some embodiments, theinternal flange 42 and the internal wall 32 can be formed from a singlepiece of a material.

In some embodiments, the rupture disk 10 consists of a rupture membrane40 and a housing 12 formed as a spool piece. In some embodiments, therupture disk 10 consists essentially of a rupture membrane 40 and ahousing 12 formed as a spool piece.

The rupture membrane 40 may be circular or non-circular when viewed froman end of the body portion 14. In some embodiments, when viewed from anend of the body portion 14, the rupture membrane 40 has a diameter, orlongest dimension of, 1-100 centimeters (cm), 1-50 cm, 1-25 cm, 1-10 cm,or 1-5 cm. In some embodiments, when viewed from an end of the bodyportion 14, the rupture membrane 40 has a diameter, or longest dimensionof, 5-100 cm, 5-50 cm, 5-25 cm, or 5-10 cm.

In some embodiments, the body portion 14 is fabricated from a firstmetal. In some embodiments, the first metal may be selected fromHastelloy®, nickel, duplex stainless steel, SAE 316L stainless steel,SAE 304 stainless steel, carbon steel, platinum, Inconel, stainlesssteel indium, aluminum, copper, lead, zinc, nickel, silver, palladium,cadmium, titanium, tungsten, tin, gold, lead, or another metal, oranother combination of these or other metals.

In some embodiments, the body portion 14 of the housing 12 is configuredas a spool piece. The rupture disk 10 can be easily installed andremoved from a system. When the rupture membrane 40 ruptures, forexample due to a sudden pressure spike, the rupture disk 10 can beeasily replaced.

Similarly, when a user needs to replace a sealing member positionedbetween the rupture disk 10 and a component to which the rupture disk 10is secured, the user can simply remove the rupture disk 10 from thecomponent, remove the sealing member, put a new sealing member in placeof the old sealing member, and re-secure the same rupture disk 10 to thecomponent.

Sealing members, which are typically made of elastomeric materials, needperiodic or event-based replacement. According to embodiments of thepresent disclosure, there is no need to replace an old rupture disk,such as the rupture disk 10, with a new rupture disk, such as therupture disk 10, simply due to the need to replace the sealing member.This allows a user to install a rupture disk 10 for the full extent ofthe rated lifetime of the rupture disk 10, which may be based on factorssuch as the ability of the rupture membrane 40 to endure cyclic loading.

There is no elastomer, such as a gasket, between the rupture membrane 40and the housing 12. In some embodiments, because there is no gasketbetween the rupture membrane 40 and the housing 12, all gaskets or othersealing members that directly engage the rupture disk 10 can be replacedwithout the need to reposition the rupture membrane 40 with respect tothe housing 12. This ensures that a seal between the rupture membrane 40and the housing 12 is maintained after replacement of the sealingmember(s).

In some embodiments, the rupture disk 10 can be incorporated into anytypical manufacturing and clean room equipment. In some embodiments, therupture disk 10 is fully suitable for cGMP (current good manufacturingpractice) processes.

The rupture disk 10 is configured to be disposed between a first andsecond component. In some embodiments, the first component may be a unitfor bioprocessing and the second component may be a reservoir. Therupture membrane 40, prior to rupture, forms a barrier between the firstcomponent and the second component. In some embodiments, the rupturedisk 10 is configured to be disposed on a first component, with anopening on the rupture disk 10 that is open to the atmosphere, so thatthe rupture disk 10, prior to rupture, forms a barrier between the firstcomponent and the atmosphere.

The rupture membrane 40 is positioned within the body portion 14. Afirst surface 46 of the rupture membrane 40 faces in a first direction,which is the direction towards the first end 30 of the body portion 14.The first surface 46 of the rupture membrane 40 is provided as a cleansurface. In some embodiments, the clean surface is free of crevices. Insome embodiments, the clean surface is configured for engaging abiological product during a biological process, such as a biologicalmanufacturing process. A second surface 48 of the rupture membrane 40faces in a second direction, which is the direction towards the secondend 34 of the body portion 14.

In some embodiments, the stepped surface 44 of the internal flange 42and the second surface 48 of the rupture membrane 40 are joined togetherby a metal-to-metal bond. In some embodiments, the metal-to-metal bondis formed in the absence of heat that results in a molten or liquidphase at the joint during a joining process. In some embodiments, thestepped surface 44 of the internal flange 42 and the second surface 48of the rupture membrane 40 are joining surfaces that are joined togetherby a cold weld. In some embodiments, the stepped surface 44 of theinternal flange 42 and the second surface 48 of the rupture membrane 40are joined together by another method. In some embodiments, the steppedsurface 44 of the internal flange 42 and the second surface 48 of therupture membrane 40 are joined together by electron-beam welding.

In FIG. 1, the rupture membrane 40 is positioned against the internalflange 42, and is cold welded to the internal flange 42. In particular,the second surface 48 of the rupture membrane 40 is secured by coldwelding to the stepped surface 44 of the internal flange 42 to form ahermetic seal between the rupture membrane 40 and the body portion 14.

In some embodiments, the rupture membrane 40 is cold welded to theinternal wall 32 of the body portion 14.

The stepped surface 44 and the rupture membrane 40 are positioned withinthe body portion 14 such that the rupture membrane 40 does not extendbeyond the first end 30 of the body portion 14 of the housing 12. Insome embodiments, as shown in FIG. 1, the rupture membrane 40 isconfigured such that it does not extend outside of the housing 12 ateither end of the body portion 14 of the housing 12.

Generally, the rupture membrane 40 is disposed to minimize a volume ofdead space 50 when the face 18 of rupture disk assembly 10 is mated withanother component, such as a bioprocessing unit. In FIG. 1, the rupturemembrane 40 is positioned such that it is entirely within the spacedefined by the internal wall 32 of the body portion 14. Moreparticularly, the rupture membrane 40 is positioned such that it isentirely within the body portion 14, but close enough to the first end30 of the body portion 14 of the housing 12 to minimize the volume ofdead space 50 defined by the first surface 46 of the rupture membrane 40and a portion of the internal wall 32 of the body portion 14 between thefirst surface 46 of the rupture membrane 40 and the first end 30 of thebody portion 14. When the rupture disk 10 is secured to a component, asurface of that component would be flush with the first face 18 of thefirst flange 16 of the housing 12.

In some embodiments, the rupture membrane 40 is positioned such that itis entirely within the body portion 14 such that the volume of the deadspace 50 defined by the first surface 46 of the rupture membrane 40 andthe portion of the internal wall 32 between the first face 46 of therupture membrane and the first end 30 of the body portion 14 that isadjacent a first component is no more than 10 cm³. In some embodiments,the volume of the dead space 50 defined by the first surface 46 of therupture membrane 40 and the portion of the internal wall 32 between thefirst face 46 of the rupture membrane 40 and the first end 30 of thebody portion 14 that is adjacent a first component is no more than 5,10, 15, 20, 30, 40, 50, 100, 200, 300, 400, or 500 cm³.

In some embodiments, the rupture membrane 40 is fabricated from a secondmetal. In some embodiments, the second metal of the rupture membrane 40is different from the first metal of the body portion 14. For example,the second metal may be selected from Hastelloy®, nickel, duplexstainless steel, SAE 316L stainless steel, SAE 304 stainless steel,carbon steel, platinum, Inconel, stainless steel indium, aluminum,copper, lead, zinc, nickel, silver, palladium, cadmium, titanium,tungsten, tin, gold, lead, or another metal, or another combination ofthese or other metals

In some embodiments, the rupture membrane 40 is scored on the firstsurface 46 and/or the second surface 48 of the rupture membrane 40. Insome embodiments, the rupture membrane 40 is configured to rupture at aspecific pressure. In some embodiments, the rupture membrane 40 can beexposed to repeated pressurization cycles before needing to be replaced.

The rupture disk 10 is configured to form a barrier that seals anaperture of a component to which the body portion 14 of the rupture disk10 is attached. In some embodiments, the rupture disk 10 forms ahermetic seal of an aperture of a component to which the body portion 14of the rupture disk 10 is attached. The rupture membrane 40, prior torupture, prevents flow of a fluid out of the aperture of the componentto which the body portion 14 of the rupture disk 10 is attached. In someembodiments, the aperture is greater than 10 mm. In some embodiments,the component is suitable for use in the production of a biologicalproduct. In some embodiments, the component comprises a chamber orreservoir or other void, and the chamber or reservoir or other void hasa linear dimension of greater than 20 centimeters.

According to an aspect of the present disclosure, FIGS. 3A-3C depict anembodiment of a rupture disk assembly shown generally at 300. Therupture disk assembly 300 includes a rupture disk housing 310 and arupture membrane carrier 312. FIG. 3A shows the rupture disk housing310. FIG. 3B shows the rupture membrane carrier 312. FIG. 3C shows therupture membrane carrier 312 secured within the rupture disk housing 310to form the assembled rupture disk assembly 300.

The rupture disk housing 310 includes a body portion 318 having aninternal wall 314 that extends from a first end 316 of the body portion318 of the rupture disk housing 310 to a second end 320 of the bodyportion 318 of the rupture disk housing 310. The internal wall 314defines an opening extending through the body portion 318 of the rupturedisk housing 310. In some embodiments, the body portion of the rupturedisk housing 310 includes an internal flange formed on the internal wall314. In some embodiments, the internal flange has a stepped surface.

The rupture membrane carrier 312 includes an outer flange portion 330that is annular and that terminates at an outer edge 332 of the rupturemembrane carrier 312. The rupture membrane carrier 312 is positionedwithin the body portion of the rupture disk housing 310 in FIG. 3C. Theouter edge 332 of the rupture membrane carrier 312 is secured to theinternal wall 314 of the rupture membrane carrier 312 to form a hermeticseal between the rupture membrane carrier 312 and the rupture diskhousing 310. In some embodiments, the outer edge 332 of the rupturemembrane carrier 312 is secured to the internal wall 314 of the rupturemembrane carrier 312 by cold welding to form a hermetic seal between therupture membrane carrier 312 and the rupture disk housing 310. In suchembodiments, the outer edge 332 includes a first joining surface and theinternal wall 314 includes a second joining surface in the cold weldingprocess.

In some embodiments, the rupture membrane carrier 312 includes a rupturemembrane carrier surface that is secured to a stepped surface on aninternal flange on the rupture disk housing 310 to form a hermetic seal,and the stepped surface and the rupture membrane are positioned withinthe body such that the rupture membrane does not extend beyond the firstend of the body.

The rupture membrane carrier 312 includes a rupture membrane portion 334that has a first rupture membrane surface 336, which is convex. Theconvex first rupture membrane surface 336 is oriented to face a fluid,such as a bioprocess fluid, which exerts a pressure in direction A onthe first rupture membrane surface 336.

A volume 340 defined by the first rupture membrane surface 336 and theportion of the internal wall 314 between the first rupture membranesurface 336 and the first end 316 of the body portion of the rupturedisk housing 310 is minimized.

The volume 340 is a clean volume, which is configured to contain abioprocess fluid during processing of the bioprocess fluid. In someembodiments, the convex first rupture membrane surface 336 is free ofcrevices, and the internal wall 314 is free of crevices.

On the opposite side of the rupture membrane portion 334 from the volume340 is a non-pressure volume 342. In typical operation, the non-pressurevolume 342 does not contact the bioprocess fluid until the rupturemembrane portion 334 ruptures.

The first rupture membrane surface 336, the internal wall 314, and thecold weld between the rupture membrane carrier 312 and the rupture diskhousing 310 provide surfaces that are free of crevices. The rupture diskassembly 300 is configured for use in bioprocessing, and in other cleanenvironments.

In some embodiments, the rupture disk assembly 10 or the rupture diskassembly 300 further includes a packaging component configured toprotect the rupture membrane 80 or the rupture membrane portion 334 fromdamage or unwanted contact with other objects. Typically, the packagingcomponent protects the pressure-side face of the rupture membrane, whichis the first surface 46 in FIG. 1 and is the first rupture membranesurface 336 in FIG. 3C. For example, the rupture disk assembly caninclude a cap, e.g., a polymeric or plastic cap, configured to fit overthe aperture of the housing nearest the rupture membrane. The cap can beconfigured to fit tightly but removably from the rupture disk assembly.The rupture disk assembly can include a second cap configured and placedso as to protect the non-pressure-side face of the rupture membrane.

According to another aspect of the present disclosure, a process systemincludes a rupture disk according to any of the embodiments describedabove and a second component. The rupture disk and the second componentare in fluid connection. In some embodiments, the process systemincludes a bioreactor. In some embodiments, the process system is abioprocess system.

In some embodiments, the second component comprises a chamber orreservoir in liquid communication with the rupture disk. In someembodiments, the chamber or reservoir vessel has a capacity or volume ofat least 0.1, 0.5, 1.0, 10, 20, 30, 40, 50, 100, 200, 500, 1,000, 1,500,10,000, 12,500, 15,00, 17,500, 20,000, 25,000 or 30,000 L. In someembodiments, the second component comprises a wall or housing, such as awall or housing defining at least a portion of the chamber or reservoir.In some embodiments, the thickness of the wall or housing is greaterthan 1, 10, 5, 10, 15, or 20 mm.

According to another aspect of the present disclosure, a method ofmaintaining a process system is provided. In some embodiments, themethod includes a step of detaching a rupture disk of any of theembodiments described above from a second component. The method thenincludes reattaching the rupture disk to a component, such as the secondcomponent from which the rupture disk was detached, thereby maintaininga process system. In some embodiments, the process system is abioprocess system. A bioprocess system may include a bioreactor orprocessing system, such as the reactors and/or components discussedherein.

With reference to FIG. 1, in an embodiment, the rupture disk assembly 10is provided on a vessel e.g., a processing tank, having a portconfigured for attachment to the rupture disk assembly 10. In anembodiment the port comprises a sanitary ferrule configured to allowattachment of the rupture disk assembly 10, e.g., by use of a sanitaryclamp.

With reference to FIG. 1, in an embodiment, the rupture disk assembly 10is disposed within the housing 12. In an embodiment, the rupture disk ispositioned deeply enough within the housing 12 that it does not protrudebeyond the housing 12. In an embodiment, the rupture disk is recessedbelow the face of the housing. In an embodiment, the rupture diskassembly 10 is positioned, e.g., to facilitate cleaning, provided it ispositioned deeply enough so as not to protrude into the space or volumebetween the pressure-side surface of the rupture disk and the housing.

The rupture disk 10 or the rupture disk 300 may be configured for usewith other components that comprise systems that have one of variousspecialized applications such as one or more of, but not limited to:sterile reception/storage of cells; automated mixing and delivery ofreagents for protein expression, production, modification (e.g.,post-translational modification) and/or secretion; automated monitoringprotein expression, production, modification, and/or secretion; cellsorting and selection, including safe waste collection; cell washing andcell collection; cell seeding on or within a proliferation substrate orscaffold; automated mixing and delivery of proliferation reagents;proliferation of cells to expand cell populations; automated monitoringof cell conditions, including detection of confluence or growth phase;controlled cell release from the proliferation substrate or scaffold;repeated proliferation steps on selected surface area sizes to increasecell numbers; cell seeding on or within culture scaffold or matrix;automatic monitoring of cell/tissue culture conditions; automaticmonitoring of protein expression or secretion; mechanical and/orbiochemical stimulation to promote proliferation; purification of theprotein and/or recovery of the protein; and storage and transportationof cells and/or protein product.

A culture vessel may be rotated or agitated within the overall devicevia control actuators. Rotation may enable the beneficial use of gravityto effect specific bioprocessing sequences such as sedimentation-basedcell seeding and fluid exchange within the bioreactor.

In embodiments, the system comprises a vessel having a housing havingone or more inlet ports and one or more outlet ports for media flow andat least one chamber defined within said bioreactor housing forreceiving cells and facilitating cell culture and protein production.The chamber may be selected from the group consisting of a cellculture/proliferation chamber and/or protein production chamber.Furthermore, the chamber houses one or more substrates and/or scaffolds.In embodiments of the disclosure, the two chambers may be providedoperably connected within the bioreactor and be operably connected.Alternatively, the two chambers may be independently operable orco-operatively operable. In still further aspects, the chambers and/orbioreactors are operably connected to provide for the exchange offluids, cells and/or tissues between the chambers and/or bioreactors.The scaffold for use in the present disclosure is selected from thegroup consisting of a porous scaffold, a porous scaffold with gradientporosity, a porous reticulate scaffold, a fibrous scaffold, a membraneencircled scaffold and combinations thereof. Funnels or similarpassageways may be provided between chambers within a bioreactor.Furthermore, one or more filters may be provided at any location withina bioreactor.

The cell culture device described herein in various embodiments is underthe control of one or more microprocessors that may be preprogrammed inorder that the user can select a specific type of environment (orsequence of environments) within the bioreactor such as cellproliferation, cell maintenance, protein production, or proteinsecretion. This eliminates operator intervention and reduces thepossibility of inadvertent contamination.

Cold Welding

Cold welding, or contact welding, is a solid-state welding process inwhich joining takes place without fusion/heating at the interface of thetwo parts to be welded. Unlike in fusion-welding processes, inembodiments, no liquid or molten phase is present when the joint or weldis formed.

Cold welding has traditionally been associated with joining metalsurfaces under ambient vacuum and/or contacting them with relativelylarge force, e.g., pressing them together at high pressure. For example,it was first recognized as a general materials phenomenon in the 1940swith the discovery that two clean, flat surfaces of similar metal wouldstrongly adhere if brought into contact under vacuum.

Cold welding methods suitable for use in, or which can be adapted foruse in, methods and devices disclosed herein are described in, e.g.,U.S. Pat. No. 8,191,756. In an embodiment a first substrate having atleast one first joint structure which comprises a first joining surface,which surface comprises a first metal; providing a second substratehaving at least one second joint structure which comprises a secondjoining surface, which surface comprises a second metal; and compressingtogether the at least one first joint structure and the at least onesecond joint structure to locally deform and shear the joining surfacesat one or more interfaces in an amount effective to form ametal-to-metal bond between the first metal and second metal of thejoining surfaces. Overlaps at the joining surfaces are effective todisplace surface contaminants and facilitate intimate contact betweenthe joining surfaces without heat input.

The first metal and second metal to be cold-welded may be the same ordifferent. They could be different alloys of the same metal. If the samemetal, the first metal and the second metal may have differentstructural morphologies, e.g., crystal structures, grain structure, etc.Non-limiting examples of suitable metal surface materials includeindium, aluminum, copper, lead, zinc, nickel, silver, palladium,cadmium, titanium, tungsten, tin, gold, or lead and combinationsthereof.

Device Components and Materials

The hermetic seal formed by affixing a rupture disk to a spool piececomprises a first substrate having at least one first joint structurewith a first joining surface and a second substrate having at least onesecond joint structure with a second joining surface, bonded at one ormore interfaces by cold welding. In embodiments, the seals arebiocompatible and suited for use with bioreactors, e.g., a bioreactor orother component disclosed in WO2014/095571.

In one embodiment, the two substrates may optionally contain or beincorporated in one or more of reservoirs, sensors, drugs, andelectronics. The substrates may comprise, silicon, glass, Pyrex® glass,stainless steel, titanium, alumina, silicon nitride, and otherbiocompatible ceramics and other metals or polymers. In one embodiment,silicon substrates allow for use of optical probes in the near-infared(NIR) to infrared (IR) spectrum. It is understood that spectroscopicmethods using light in the visible, UV or other wavelengths may bepossible by an appropriate selection of substrate material. In addition,the substrate may comprise polymers with high enough Young's Modulus andyield stress to cause high shear during cold welding.

The joint structures have joining surfaces (also called “shear layers”or “bonding surfaces”) which are preferably metal and optionally maybond to other joining surfaces. In an alternate embodiment, described infurther detail below, the joining surface may be a compliant polymer.Metals with a suitably low plastic deformation stress are used as ajoining surface. Suitability can be determined by one skilled in theart, for example, based on the particular joint geometry and the amountof force that can reasonably be applied to form the joint. In addition,metals that do not have a surface oxide or have a high relative oxide toparent metal hardness are preferable for use as a joining surface. SeeTylecote, “Investigations on Pressure Welding” British Welding J. (March1954) and Mohamed, et al., “Mechanism of Solid State Pressure Welding”Welding Research Supplement, pp. 302-10 (September 1975). Representativeexamples of suitable metals (and their alloys) include gold (Au), indium(In), aluminum (Al), copper (Cu), lead (Pb), zinc (Zn), nickel (Ni),silver (Ag), platinum (Pt), palladium (Pd), and cadmium (Cd).Representative examples of joining surface metals preferred forbiocompatibility include gold and platinum.

The first joining surface may or may not be comprised of the samematerial as the second joining surface with which the first joiningsurface will form the hermetic seal. For example, the joining surfacesmay be comprised of dissimilar metals or different alloys of the sameparent metal. For example, the first joining surface may be gold whilethe second is platinum. In one embodiment, the joining surfaces arecomprised of the same material with a different structural morphology.For instance, a first joining surface may be annealed to reduce theyield stress through the normal annealing mechanisms of recovery,recrystallization, and grain growth, while the second joining surfacemay be deposited in such a way that the grain size is small, thusincreasing the yield stress.

The joining surfaces may comprise the same or a different material thanthe joint structures. This allows greater freedom in the fabricationmethod of the joint as well as more design control over the extent andlocation of plastic deformation. The joint structure may be comprised ofa single material or a combination of materials.

Methods of Making a Hermetic Seal

In embodiments, the hermetic seals are made by compression and coldwelding. In one embodiment, two substrates are hermetically sealedtogether by providing a first substrate having at least one first jointstructure which comprises a first joining surface which is a metal,providing a second substrate having at least one second joint structurewhich comprises a second joining surface which is a metal, compressingtogether at least one first joint structure and at least one secondjoint structure to locally deform and shear the metal surfaces at one ormore interfaces in an amount effective to form a continuousmetal-to-metal bond between the joining surfaces at the one or moreinterfaces.

In some embodiments, ultrasonic energy may be introduced to the hermeticseal joint during the bonding process. While not being bound to anyparticular mechanism of action, it is believed that the ultrasonicenergy may improve the hermetic seal by causing metal-to-metalinter-diffusion by scrubbing the contaminants out of the joiningsurfaces and deforming the surface asperities so there is intimatecontact at the bonding interface.

In other embodiments where the bonding mechanism is not purely coldwelding, a pulse of heat or a small increase in temperature may aid inmetal bonding by increasing diffusion and lowering the metal's yieldstress. For example, induction heating could be used to locally heat thejoining surface metals. If other metals are present in the device andare non-magnetic, the joining metals can be selectively heated byincorporating a magnetic material under the joining surfaces.

Representative examples of magnetic materials include nickel, iron,cobalt, and combinations thereof. Alternatively, the joint structuregeometry may be designed to selectively couple a magnetic field of agiven frequency. (See Cao et al., “Selective and localized bonding usinginduction heating”, Solid-State Sensor, Actuator and MicrosystemsWorkshop, Hilton Head Island, S.C., Jun. 2-6, 2002.)

Generally, the ambient environment may be displaced with forming gas,nitrogen, vacuum, or some other condition which would minimize the rateof oxidation and contamination of the joining surfaces as the hermeticbond is formed.

Reactors/Components

The rupture disk assemblies disclosed herein can be used with abioreactor or processing vessel or tank, or, more generally with anyfeed source. The devices, facilities and methods described herein aresuitable for culturing any desired cell line including prokaryoticand/or eukaryotic cell lines. Further, in embodiments, the devices,facilities and methods are suitable for culturing suspension cells oranchorage-dependent (adherent) cells and are suitable for productionoperations configured for production of pharmaceutical andbiopharmaceutical products—such as polypeptide products, nucleic acidproducts (for example DNA or RNA), or cells and/or viruses such as thoseused in cellular and/or viral therapies.

In embodiments, the cells express or produce a product, such as arecombinant therapeutic or diagnostic product. As described in moredetail below, examples of products produced by cells include, but arenot limited to, antibody molecules (e.g., monoclonal antibodies,bispecific antibodies), antibody mimetics (polypeptide molecules thatbind specifically to antigens but that are not structurally related toantibodies such as e.g. DARPins, affibodies, adnectins, or IgNARs),fusion proteins (e.g., Fc fusion proteins, chimeric cytokines), otherrecombinant proteins (e.g., glycosylated proteins, enzymes, hormones),viral therapeutics (e.g., anti-cancer oncolytic viruses, viral vectorsfor gene therapy and viral immunotherapy), cell therapeutics (e.g.,pluripotent stem cells, mesenchymal stem cells and adult stem cells),vaccines or lipid-encapsulated particles (e.g., exosomes, virus-likeparticles), RNA (such as e.g. siRNA) or DNA (such as e.g. plasmid DNA),antibiotics or amino acids. In embodiments, the devices, facilities andmethods can be used for producing biosimilars.

As mentioned, in embodiments, devices, facilities and methods allow forthe production of eukaryotic cells, e.g., mammalian cells or lowereukaryotic cells such as for example yeast cells or filamentous fungicells, or prokaryotic cells such as Gram-positive or Gram-negative cellsand/or products of the eukaryotic or prokaryotic cells, e.g., proteins,peptides, antibiotics, amino acids, nucleic acids (such as DNA or RNA),synthesised by the eukaryotic cells in a large-scale manner. Unlessstated otherwise herein, the devices, facilities, and methods caninclude any desired volume or production capacity including but notlimited to bench-scale, pilot-scale, and full production scalecapacities.

Moreover and unless stated otherwise herein, the devices, facilities,and methods can include any suitable reactor(s) including but notlimited to stirred tank, airlift, fiber, microfiber, hollow fiber,ceramic matrix, fluidized bed, fixed bed, and/or spouted bedbioreactors. As used herein, “reactor” can include a fermentor orfermentation unit, or any other reaction vessel and the term “reactor”is used interchangeably with “fermentor.” For example, in some aspects,a bioreactor unit can perform one or more, or all, of the following:feeding of nutrients and/or carbon sources, injection of suitable gas(e.g., oxygen), inlet and outlet flow of fermentation or cell culturemedium, separation of gas and liquid phases, maintenance of temperature,maintenance of oxygen and CO2 levels, maintenance of pH level, agitation(e.g., stirring), and/or cleaning/sterilizing. Example reactor units,such as a fermentation unit, may contain multiple reactors within theunit, for example the unit can have 1, 2, 3, 4, 5, 10, 15, 20, 25, 30,35, 40, 45, 50, 60, 70, 80, 90, or 100, or more bioreactors in each unitand/or a facility may contain multiple units having a single or multiplereactors within the facility. In various embodiments, the bioreactor canbe suitable for batch, semi fed-batch, fed-batch, perfusion, and/or acontinuous fermentation processes. Any suitable reactor diameter can beused. In embodiments, the bioreactor can have a volume between about 100mL and about 50,000 L. Non-limiting examples include a volume of 100 mL,250 mL, 500 mL, 750 mL, 1 liter, 2 liters, 3 liters, 4 liters, 5 liters,6 liters, 7 liters, 8 liters, 9 liters, 10 liters, 15 liters, 20 liters,25 liters, 30 liters, 40 liters, 50 liters, 60 liters, 70 liters, 80liters, 90 liters, 100 liters, 150 liters, 200 liters, 250 liters, 300liters, 350 liters, 400 liters, 450 liters, 500 liters, 550 liters, 600liters, 650 liters, 700 liters, 750 liters, 800 liters, 850 liters, 900liters, 950 liters, 1000 liters, 1500 liters, 2000 liters, 2500 liters,3000 liters, 3500 liters, 4000 liters, 4500 liters, 5000 liters, 6000liters, 7000 liters, 8000 liters, 9000 liters, 10,000 liters, 15,000liters, 20,000 liters, and/or 50,000 liters. Additionally, suitablereactors can be multi-use, single-use, disposable, or non-disposable andcan be formed of any suitable material including metal alloys such asstainless steel (e.g., 316L or any other suitable stainless steel) andInconel, plastics, and/or glass.

In embodiments and unless stated otherwise herein, the devices,facilities, and methods described herein can also include any suitableunit operation and/or equipment not otherwise mentioned, such asoperations and/or equipment for separation, purification, and isolationof such products. Any suitable facility and environment can be used,such as traditional stick-built facilities, modular, mobile andtemporary facilities, or any other suitable construction, facility,and/or layout. For example, in some embodiments modular clean-rooms canbe used. Additionally and unless otherwise stated, the devices, systems,and methods described herein can be housed and/or performed in a singlelocation or facility or alternatively be housed and/or performed atseparate or multiple locations and/or facilities.

By way of non-limiting examples and without limitation, U.S. PublicationNos. 2013/0280797; 2012/0077429;2009/0305626; and U.S. Pat. Nos.8,298,054; 7,629,167; and 5,656,491, which are hereby incorporated byreference in their entirety, describe example facilities, equipment,and/or systems that may be suitable.

In embodiments, the cells are eukaryotic cells, e.g., mammalian cells.The mammalian cells can be for example human or rodent or bovine celllines or cell strains. Examples of such cells, cell lines or cellstrains are e.g. mouse myeloma (NSO)-cell lines, Chinese hamster ovary(CHO)-cell lines, HT1080, H9, HepG2, MCF7, MDBK Jurkat, NIH3T3, PC12,BHK (baby hamster kidney cell), VERO, SP2/0, YB2/0, YO, C127, L cell,COS, e.g., COS 1 and COS7, QC1-3,HEK-293, VERO, PER.C6, HeLA, EB1, EB2,EB3, oncolytic or hybridoma-cell lines. Preferably the mammalian cellsare CHO-cell lines. In one embodiment, the cell is a CHO cell. In oneembodiment, the cell is a CHO-K1 cell, a CHO-K1 SV cell, a DG44 CHOcell, a DUXB11 CHO cell, a CHOS, a CHO GS knock-out cell, a CHO FUT8 GSknock-out cell, a CHOZN, or a CHO-derived cell. The CHO GS knock-outcell (e.g., GSKO cell) is, for example, a CHO-K1 SV GS knockout cell.The CHO FUT8 knockout cell is, for example, the Potelligent® CHOK1 SV(Lonza Biologics, Inc.). Eukaryotic cells can also be avian cells, celllines or cell strains, such as for example, EBx® cells, EB14, EB24,EB26, EB66, or EBv13.

In one embodiment, the eukaryotic cells are stem cells. The stem cellscan be, for example, pluripotent stem cells, including embryonic stemcells (ESCs), adult stem cells, induced pluripotent stem cells (iPSCs),tissue specific stem cells (e.g., hematopoietic stem cells) andmesenchymal stem cells (MSCs).

In one embodiment, the cell is a differentiated form of any of the cellsdescribed herein. In one embodiment, the cell is a cell derived from anyprimary cell in culture.

In embodiments, the cell is a hepatocyte such as a human hepatocyte,animal hepatocyte, or a non-parenchymal cell. For example, the cell canbe a plateable metabolism qualified human hepatocyte, a plateableinduction qualified human hepatocyte, plateable Qualyst TransporterCertified™ human hepatocyte, suspension qualified human hepatocyte(including 10-donor and 20-donor pooled hepatocytes), human hepatickupffer cells, human hepatic stellate cells, dog hepatocytes (includingsingle and pooled Beagle hepatocytes), mouse hepatocytes (including CD-1and C57BI/6 hepatocytes), rat hepatocytes (including Sprague-Dawley,Wistar Han, and Wistar hepatocytes), monkey hepatocytes (includingCynomolgus or Rhesus monkey hepatocytes), cat hepatocytes (includingDomestic Shorthair hepatocytes), and rabbit hepatocytes (including NewZealand White hepatocytes). Example hepatocytes are commerciallyavailable from Triangle Research Labs, LLC, 6 Davis Drive ResearchTriangle Park, North Carolina, USA 27709.

In one embodiment, the eukaryotic cell is a lower eukaryotic cell suchas e.g. a yeast cell (e.g., Pichia genus (e.g. Pichia pastoris, Pichiamethanolica, Pichia kluyveri, and Pichia angusta), Komagataella genus(e.g. Komagataella pastoris, Komagataella pseudopastoris or Komagataellaphaffii), Saccharomyces genus (e.g. Saccharomyces cerevisae, cerevisiae,Saccharomyces kluyveri, Saccharomyces uvarum), Kluyveromyces genus (e.g.Kluyveromyces lactis, Kluyveromyces marxianus), the Candida genus (e.g.Candida utilis, Candida cacaoi, Candida boidinii,), the Geotrichum genus(e.g. Geotrichum fermentans), Hansenula polymorpha, Yarrowia lipolytica,or Schizosaccharomyces pombe. Preferred is the species Pichia pastoris.Examples for Pichia pastoris strains are X33, GS115, KM71, KM71H; andCBS7435.

In one embodiment, the eukaryotic cell is a fungal cell (e.g.Aspergillus (such as A. niger, A. fumigatus, A. orzyae, A. nidula),Acremonium (such as A. thermophilum), Chaetomium (such as C.thermophilum), Chrysosporium (such as C. thermophile), Cordyceps (suchas C. militaris), Corynascus, Ctenomyces, Fusarium (such as F.oxysporum), Glomerella (such as G. graminicola), Hypocrea (such as H.jecorina), Magnaporthe (such as M. orzyae), Myceliophthora (such as M.thermophile), Nectria (such as N. heamatococca), Neurospora (such as N.crassa), Penicillium, Sporotrichum (such as S. thermophile), Thielavia(such as T. terrestris, T. heterothallica), Trichoderma (such as T.reesei), or Verticillium (such as V. dahlia)).

In one embodiment, the eukaryotic cell is an insect cell (e.g., Sf9,Mimic™ Sf9, Sf21, High Five™ (BT1-TN-5B1-4), or BT1-Ea88 cells), analgae cell (e.g., of the genus Amphora, Bacillariophyceae, Dunaliella,Chlorella, Chlamydomonas, Cyanophyta (cyanobacteria), Nannochloropsis,Spirulina, or Ochromonas), or a plant cell (e.g., cells frommonocotyledonous plants (e.g., maize, rice, wheat, or Setaria), or froma dicotyledonous plants (e.g., cassava, potato, soybean, tomato,tobacco, alfalfa, Physcomitrella patens or Arabidopsis).

In one embodiment, the cell is a bacterial or prokaryotic cell.

In embodiments, the prokaryotic cell is a Gram-positive cells such asBacillus, Streptomyces Streptococcus, Staphylococcus or Lactobacillus.Bacillus that can be used is, e.g. the B. subtilis, B.amyloliquefaciens, B. licheniformis, B. natto, or B. megaterium. Inembodiments, the cell is B. subtilis, such as B. subtilis 3NA and B.subtilis 168. Bacillus is obtainable from, e.g., the Bacillus GeneticStock Center, Biological Sciences 556, 484 West 12^(th) Avenue, ColumbusOhio 43210-1214.

In one embodiment, the prokaryotic cell is a Gram-negative cell, such asSalmonella spp. or Escherichia coli, such as e.g., TG1, TG2, W3110, DH1,DHB4, DH5a, HMS 174, HMS174 (DE3), NM533, C600, HB101, JM109, MC4100,XL1-Blue and Origami, as well as those derived from E. coli B-strains,such as for example BL-21 or BL21 (DE3), all of which are commerciallyavailable.

Suitable host cells are commercially available, for example, fromculture collections such as the DSMZ (Deutsche Sammlung vonMikroorganismen and Zellkulturen GmbH, Braunschweig, Germany) or theAmerican Type Culture Collection (ATCC).

In embodiments, the cultured cells are used to produce proteins e.g.,antibodies, e.g., monoclonal antibodies, and/or recombinant proteins,for therapeutic use. In embodiments, the cultured cells producepeptides, amino acids, fatty acids or other useful biochemicalintermediates or metabolites. For example, in embodiments, moleculeshaving a molecular weight of about 4000 daltons to greater than about140,000 daltons can be produced. In embodiments, these molecules canhave a range of complexity and can include posttranslationalmodifications including glycosylation.

In embodiments, the protein is, e.g., BOTOX, Myobloc, Neurobloc, Dysport(or other serotypes of botulinum neurotoxins), alglucosidase alpha,daptomycin, YH-16, choriogonadotropin alpha, filgrastim, cetrorelix,interleukin-2, aldesleukin, teceleulin, denileukin diftitox, interferonalpha-n3 (injection), interferon alpha-n1, DL-8234, interferon, Suntory(gamma-1a), interferon gamma, thymosin alpha 1, tasonermin, DigiFab,ViperaTAb, EchiTAb, CroFab, nesiritide, abatacept, alefacept, Rebif,eptoterminalfa, teriparatide (osteoporosis), calcitonin injectable (bonedisease), calcitonin (nasal, osteoporosis), etanercept, hemoglobinglutamer 250 (bovine), drotrecogin alpha, collagenase, carperitide,recombinant human epidermal growth factor (topical gel, wound healing),DWP401, darbepoetin alpha, epoetin omega, epoetin beta, epoetin alpha,desirudin, lepirudin, bivalirudin, nonacog alpha, Mononine, eptacogalpha (activated), recombinant Factor VIII+VWF, Recombinate, recombinantFactor VIII, Factor VIII (recombinant), Alphnmate, octocog alpha, FactorVIII, palifermin,Indikinase, tenecteplase, alteplase, pamiteplase,reteplase, nateplase, monteplase, follitropin alpha, rFSH, hpFSH,micafungin, pegfilgrastim, lenograstim, nartograstim, sermorelin,glucagon, exenatide, pramlintide, iniglucerase, galsulfase, Leucotropin,molgramostim, triptorelin acetate, histrelin (subcutaneous implant,Hydron), deslorelin, histrelin, nafarelin, leuprolide sustained releasedepot (ATRIGEL), leuprolide implant (DUROS), goserelin, Eutropin, KP-102program, somatropin, mecasermin (growth failure), enlfavirtide,Org-33408, insulin glargine, insulin glulisine, insulin (inhaled),insulin lispro, insulin deternir, insulin (buccal, RapidMist),mecasermin rinfabate, anakinra, celmoleukin, 99 mTc-apcitide injection,myelopid, Betaseron, glatiramer acetate, Gepon, sargramostim,oprelvekin, human leukocyte-derived alpha interferons, Bilive, insulin(recombinant), recombinant human insulin, insulin aspart, mecasenin,Roferon-A, interferon-alpha 2, Alfaferone, interferon alfacon-1,interferon alpha, Avonex' recombinant human luteinizing hormone, dornasealpha, trafermin, ziconotide, taltirelin, diboterminalfa, atosiban,becaplermin, eptifibatide, Zemaira, CTC-111, Shanvac-B, HPV vaccine(quadrivalent), octreotide, lanreotide, ancestirn, agalsidase beta,agalsidase alpha, laronidase, prezatide copper acetate (topical gel),rasburicase, ranibizumab, Actimmune, PEG-Intron, Tricomin, recombinanthouse dust mite allergy desensitization injection, recombinant humanparathyroid hormone (PTH) 1-84 (sc, osteoporosis), epoetin delta,transgenic antithrombin III, Granditropin, Vitrase, recombinant insulin,interferon-alpha (oral lozenge), GEM-21S, vapreotide, idursulfase,omnapatrilat, recombinant serum albumin, certolizumab pegol,glucarpidase, human recombinant C1 esterase inhibitor (angioedema),lanoteplase, recombinant human growth hormone, enfuvirtide (needle-freeinjection, Biojector 2000), VGV-1, interferon (alpha), lucinactant,aviptadil (inhaled, pulmonary disease), icatibant, ecallantide,omiganan, Aurograb, pexigananacetate, ADI-PEG-20, LDI-200, degarelix,cintredelinbesudotox, Favld, MDX-1379, ISAtx-247, liraglutide,teriparatide (osteoporosis), tifacogin, AA4500, T4N5 liposome lotion,catumaxomab, DWP413, ART-123, Chrysalin, desmoteplase, amediplase,corifollitropinalpha, TH-9507, teduglutide, Diamyd, DWP-412, growthhormone (sustained release injection), recombinant G-CSF, insulin(inhaled, AIR), insulin (inhaled, Technosphere), insulin (inhaled,AERx), RGN-303, DiaPep277, interferon beta (hepatitis C viral infection(HCV)), interferon alpha-n3 (oral), belatacept, transdermal insulinpatches, AMG-531, MBP-8298, Xerecept, opebacan, AIDSVAX, GV-1001,LymphoScan, ranpirnase, Lipoxysan, lusupultide, MP52(beta-tricalciumphosphate carrier, bone regeneration), melanoma vaccine,sipuleucel-T, CTP-37, Insegia, vitespen, human thrombin (frozen,surgical bleeding), thrombin, TransMlD, alfimeprase, Puricase, terlipressin (intravenous, hepatorenal syndrome), EUR-1008M, recombinant FGF-I(injectable, vascular disease), BDM-E, rotigaptide, ETC-216, P-113,MBI-594AN, duramycin (inhaled, cystic fibrosis), SCV-07, OPI-45,Endostatin, Angiostatin, ABT-510, Bowman Birk Inhibitor Concentrate,XMP-629, 99 mTc-Hynic-Annexin V, kahalalide F, CTCE-9908, teverelix(extended release), ozarelix, rornidepsin, BAY-504798, interleukin4,PRX-321, Pepscan, iboctadekin, rhlactoferrin, TRU-015, IL-21, ATN-161,cilengitide, Albuferon, Biphasix, IRX-2, omega interferon, PCK-3145,CAP-232, pasireotide, huN901-DMI, ovarian cancer immunotherapeuticvaccine, SB-249553, Oncovax-CL, OncoVax-P, BLP-25, CerVax-16,multi-epitope peptide melanoma vaccine (MART-1, gp100, tyrosinase),nemifitide, rAAT (inhaled), rAAT (dermatological), CGRP (inhaled,asthma), pegsunercept, thymosinbeta4, plitidepsin, GTP-200, ramoplanin,GRASPA, OBI-1, AC-100, salmon calcitonin (oral, eligen), calcitonin(oral, osteoporosis), examorelin, capromorelin, Cardeva, velafermin,131I-TM-601, KK-220, T-10, ularitide, depelestat, hematide, Chrysalin(topical), rNAPc2, recombinant Factor V111 (PEGylated liposomal), bFGF,PEGylated recombinant staphylokinase variant, V-10153, SonoLysisProlyse, NeuroVax, CZEN-002, islet cell neogenesis therapy, rGLP-1,BIM-51077, LY-548806, exenatide (controlled release, Medisorb),AVE-0010, GA-GCB, avorelin, ACM-9604, linaclotid eacetate, CETi-1,Hemospan, VAL (injectable), fast-acting insulin (injectable, Viadel),intranasal insulin, insulin (inhaled), insulin (oral, eligen),recombinant methionyl human leptin, pitrakinra subcutancous injection,eczema), pitrakinra (inhaled dry powder, asthma), Multikine, RG-1068,MM-093, NBI-6024, AT-001, PI-0824, Org-39141, Cpn10 (autoimmunediseases/inflammation), talactoferrin (topical), rEV-131 (ophthalmic),rEV-131 (respiratory disease), oral recombinant human insulin(diabetes), RPI-78M, oprelvekin (oral), CYT-99007 CTLA4-Ig, DTY-001,valategrast, interferon alpha-n3 (topical), IRX-3, RDP-58, Tauferon,bile salt stimulated lipase, Merispase, alaline phosphatase, EP-2104R,Melanotan-II, bremelanotide, ATL-104, recombinant human microplasmin,AX-200, SEMAX, ACV-1, Xen-2174, CJC-1008, dynorphin A, SI-6603, LABGHRH, AER-002, BGC-728, malaria vaccine (virosomes, PeviPRO), ALTU-135,parvovirus B19 vaccine, influenza vaccine (recombinant neuraminidase),malaria/HBV vaccine, anthrax vaccine, Vacc-5q, Vacc-4x, HIV vaccine(oral), HPV vaccine, Tat Toxoid, YSPSL, CHS-13340, PTH(1-34) liposomalcream (Novasome), Ostabolin-C, PTH analog (topical, psoriasis),MBRI-93.02, MTB72F vaccine (tuberculosis), MVA-Ag85A vaccine(tuberculosis), FARA04, BA-210, recombinant plague FIV vaccine, AG-702,OxSODrol, rBetV1, Der-p1/Der-p2/Der-p7 allergen-targeting vaccine (dustmite allergy), PR1 peptide antigen (leukemia), mutant ras vaccine,HPV-16 E7 lipopeptide vaccine, labyrinthin vaccine (adenocarcinoma), CMLvaccine, WT1-peptide vaccine (cancer), IDD-5, CDX-110, Pentrys, Norelin,CytoFab, P-9808, VT-111, icrocaptide, telbermin (dermatological,diabetic foot ulcer), rupintrivir, reticulose, rGRF, HA,alpha-galactosidase A, ACE-011, ALTU-140, CGX-1160, angiotensintherapeutic vaccine, D-4F, ETC-642, APP-018, rhMBL, SCV-07 (oral,tuberculosis), DRF-7295, ABT-828, ErbB2-specific immunotoxin(anticancer), DT3SSIL-3, TST-10088, PRO-1762, Combotox,cholecystokinin-B/gastrin-receptor binding peptides, 111In-hEGF, AE-37,trasnizumab-DM1, Antagonist G, IL-12 (recombinant), PM-02734, IMP-321,rhIGF-BP3, BLX-883, CUV-1647 (topical), L-19 basedradioimmunotherapeutics (cancer), Re-188-P-2045, AMG-386, DC/1540/KLHvaccine (cancer), VX-001, AVE-9633, AC-9301, NY-ESO-1 vaccine(peptides), NA17.A2 peptides, melanoma vaccine (pulsed antigentherapeutic), prostate cancer vaccine, CBP-501, recombinant humanlactoferrin (dry eye), FX-06, AP-214, WAP-8294A (injectable), ACP-HIP,SUN-11031, peptide YY [3-36] (obesity, intranasal), FGLL, atacicept,BR3-Fc, BN-003, BA-058, human parathyroid hormone 1-34 (nasal,osteoporosis), F-18-CCR1, AT-1100 (celiac disease/diabetes), JPD-003,PTH(7-34) liposomal cream (Novasome), duramycin (ophthalmic, dry eye),CAB-2, CTCE-0214, GlycoPEGylated erythropoietin, EPO-Fc, CNTO-528,AMG-114, JR-013, Factor XIII, aminocandin, PN-951, 716155, SUN-E7001,TH-0318, BAY-73-7977, teverelix (immediate release), EP-51216, hGH(controlled release, Biosphere), OGP-I, sifuvirtide, TV4710, ALG-889,Org-41259, rhCC10, F-991, thymopentin (pulmonary diseases), r(m)CRP,hepatoselective insulin, subalin, L19-IL-2 fusion protein, elafin,NMK-150, ALTU-139, EN-122004, rhTPO, thrombopoietin receptor agonist(thrombocytopenic disorders), AL-108, AL-208, nerve growth factorantagonists (pain), SLV-317, CGX-1007, INNO-105, oral teriparatide(eligen), GEM-OS1, AC-162352, PRX-302, LFn-p24 fusion vaccine(Therapore), EP-1043, S pneumoniae pediatric vaccine, malaria vaccine,Neisseria meningitidis Group B vaccine, neonatal group B streptococcalvaccine, anthrax vaccine, HCV vaccine (gpE1+gpE2+MF-59), otitis mediatherapy, HCV vaccine (core antigen+ISCOMATRIX), hPTH(1-34) (transdermal,ViaDerm), 768974, SYN-101, PGN-0052, aviscumnine, BIM-23190,tuberculosis vaccine, multi-epitope tyrosinase peptide, cancer vaccine,enkastim, APC-8024, GI-5005, ACC-001, TTS-CD3, vascular-targeted TNF(solid tumors), desmopressin (buccal controlled-release), onercept, andTP-9201.

In some embodiments, the polypeptide is adalimumab (HUMIRA), infliximab(REMICADE™), rituximab (RITUXANTM/MAB THERA™) etanercept (ENBREL™)bevacizumab (AVASTIN™), trastuzumab (HERCEPTIN™), pegrilgrastim(NEULASTA™), or any other suitable polypeptide including biosimilars andbiobetters.

Other suitable polypeptides are those listed below and in Table1(adapted from US2016/0097074):

Protein Product Reference Listed Drug interferon gamma-1b Actimmune ®alteplase; tissue plasminogen activator Activase ®/Cathflo ® Recombinantantihemophilic factor Advate human albumin Albutein ® LaronidaseAldurazyme ® Interferon alfa-N3, human leukocyte derived Alferon N ®human antihemophilic factor Alphanate ® virus-filtered human coagulationfactor IX AlphaNine ® SD Alefacept; recombinant, dimeric fusion proteinAmevive ® LFA3-Ig Bivalirudin Angiomax ® darbepoetin alfa Aranesp ™Bevacizumab Avastin ™ interferon beta-1a; recombinant Avonex ®coagulation factor IX BeneFix ™ Interferon beta-1b Betaseron ®Tositumomab BEXXAR ® antihemophilic factor Bioclate ™ human growthhormone BioTropin ™ botulinum toxin type A BOTOX ® Alemtuzumab Campath ®acritumomab; technetium-99 labeled CEA-Scan ® alglucerase; modified formof beta- Ceredase ® glucocerebrosidase imiglucerase; recombinant form ofbeta- Cerezyme ® glucocerebrosidase crotalidae polyvalent immune Fab,ovine CroFab ™ digoxin immune fab [ovine] DigiFab ™ Rasburicase Elitek ®Etanercept ENBREL ® epoietin alfa Epogen ® Cetuximab Erbitux ™algasidase beta Fabrazyme ® Urofollitropin Fertinex ™ follitropin betaFollistim ™ Teriparatide FORTEO ® human somatropin GenoTropin ® GlucagonGlucaGen ® follitropin alfa Gonal-F ® antihemophilic factor Helixate ®Antihemophilic Factor; Factor XIII HEMOFIL adefovir dipivoxil Hepsera ™Trastuzumab Herceptin ® Insulin Humalog ® antihemophilic factor/vonWillebrand factor Humate-P ® complex-human Somatotropin Humatrope ®Adalimumab HUMIRA ™ human insulin Humulin ® recombinant humanhyaluronidase Hylenex ™ interferon alfacon-1 Infergen ® EptifibatideIntegrilin ™ alpha-interferon Intron A ® Palifermin Kepivance AnakinraKineret ™ antihemophilic factor Kogenate ® FS insulin glargine Lantus ®granulocyte macrophage colony-stimulating Leukine ®/Leukine ® Liquidfactor lutropin alfa for injection Luveris OspA lipoprotein LYMErix ™Ranibizumab LUCENTIS ® gemtuzumab ozogamicin Mylotarg ™ GalsulfaseNaglazyme ™ Nesiritide Natrecor ® Pegfilgrastim Neulasta ™ OprelvekinNeumega ® Filgrastim Neupogen ® Fanolesomab NeutroSpec ™ (formerlyLeuTech ®) somatropin [rDNA] Norditropin ®/Norditropin Nordiflex ®Mitoxantrone Novantrone ® insulin; zinc suspension; Novolin L ® insulin;isophane suspension Novolin N ® insulin, regular; Novolin R ® InsulinNovolin ® coagulation factor VIIa NovoSeven ® Somatropin Nutropin ®immunoglobulin intravenous Octagam ® PEG-L-asparaginase Oncaspar ®abatacept, fully human soluable fusion protein Orencia ™ muromomab-CD3Orthoclone OKT3 ® high-molecular weight hyaluronan Orthovisc ® humanchorionic gonadotropin Ovidrel ® live attenuated BacillusCalmette-Guerin Pacis ® peginterferon alfa-2a Pegasys ® pegylatedversion of interferon alfa-2b PEG-Intron ™ Abarelix (injectablesuspension); gonadotropin-releasing Plenaxis ™ hormone Antagonistepoietin alfa Procrit ® Aldesleukin Proleukin, IL-2 ® SomatremProtropin ® dornase alfa Pulmozyme ® Efalizumab; selective, reversibleT-cell blocker RAPTIVA ™ combination of ribavirin and alpha interferonRebetron ™ Interferon beta 1a Rebif ® antihemophilic factorRecombinate ® rAHF/ antihemophilic factor ReFacto ® Lepirudin Refludan ®Infliximab REMICADE ® Abciximab ReoPro ™ Reteplase Retavase ™ RituximaRituxan ™ interferon alfa-2^(a) Roferon-A ® Somatropin Saizen ®synthetic porcine secretin SecreFlo ™ Basiliximab Simulect ® EculizumabSOLIRIS (R) Pegvisomant SOMAVERT ® Palivizumab; recombinantly produced,humanized mAb Synagis ™ thyrotropin alfa Thyrogen ® TenecteplaseTNKase ™ Natalizumab TYSABRI ® human immune globulin intravenous 5% and10% solutions Venoglobulin-S ® interferon alfa-n1, lymphoblastoidWellferon ® drotrecogin alfa Xigris ™ Omalizumab; recombinantDNA-derived humanized monoclonal Xolair ® antibody targetingimmunoglobulin-E Daclizumab Zenapax ® ibritumomab tiuxetan Zevalin ™Somatotropin Zorbtive ™ (Serostim ®)

In embodiments, the polypeptide is a hormone, blood clotting/coagulationfactor, cytokine/growth factor, antibody molelcule, fusion protein,protein vaccine, or peptide as shown in Table 2, below.

TABLE 2 Exemplary Products Therapeutic Product type Product Trade NameHormone Erythropoietin, Epoein-α Epogen, Procrit Darbepoetin-α AranespGrowth hormone (GH), Genotropin, Humatrope, Norditropin, somatotropinNovIVitropin, Nutropin, Omnitrope, Protropin, Siazen, Serostim,Valtropin Gonal-F, Follistim Human follicle-stimulating Ovidrel hormone(FSH) Human chorionic Luveris gonadotropin GlcaGen Lutropin-α GerefGlucagon ChiRhoStim (human peptide), SecreFlo Growth hormone releasing(porcine peptide) hormone (GHRH) Thyrogen Secretin Thyroid stimulatinghormone (TSH), thyrotropin Blood Factor VIIa NovoSevenClotting/Coagulation Factor VIII Bioclate, Helixate, Kogenate, FactorsRecombinate, ReFacto Factor IX Antithrombin III (AT-III) Benefix ProteinC concentrate Thrombate III Ceprotin Cytokine/Growth Type Ialpha-interferon Infergen factor Interferon-αn3 (IFNαn3) Alferon NInterferon-β1a (rIFN- β) Avonex, Rebif Interferon-β1b (rIFN- β)Betaseron Interferon-γ1b (IFN γ) Actimmune Aldesleukin (interleukinProleukin 2(IL2), epidermal theymocyte activating factor; ETAFPalifermin (keratinocyte Kepivance growth factor; KGF) RegranexBecaplemin (platelet- Anril, Kineret derived growth factor; PDGF)Anakinra (recombinant IL1 antagonist) Antibody molecules Bevacizumab(VEGFA Avastin mAb) Erbitux Cetuximab (EGFR mAb) Vectibix Panitumumab(EGFR mAb) Campath Alemtuzumab (CD52 mAb) Rituxan Rituximab (CD20chimeric Herceptin Ab) Orencia Trastuzumab (HER2/Neu Humira mAb) EnbrelAbatacept (CTLA Ab/Fc Remicade fusion) Adalimumab (TNFα mAb) AmeviveEtanercept (TNF Raptiva receptor/Fc fusion) Tysabri Infliximab (TNFαchimeric Soliris mAb) Alefacept (CD2 fusion Orthoclone, OKT3 protein)Efalizumab (CD11a mAb) Natalizumab (integrin α4 subunit mAb) Eculizumab(C5mAb) Muromonab-CD3 Other: Insulin Humulin, Novolin Fusion Hepatitis Bsurface antigen Engerix, Recombivax HB proteins/Protein (HBsAg)vaccines/Peptides HPV vaccine Gardasil OspA LYMErix Anti-Rhesus(Rh)Rhophylac immunoglobulin G Fuzeon Enfuvirtide Spider silk, e.g., fibrionQMONOS

In embodiments, the protein is multispecific protein, e.g., a bispecificantibody as shown in Table 3.

TABLE 3 Bispecific Formats Name (other names, Proposed Diseases (orsponsoring BsAb mechanisms of Development healthy organizations) formatTargets action stages volunteers) Catumaxomab BsIgG: CD3, Retargeting ofT Approved in Malignant ascites (Removab ®, Triomab EpCAM cells totumor, Fc EU in EpCAM Fresenius Biotech, mediated effector positivetumors Trion Pharma, functions Neopharm) Ertumaxomab BsIgG: CD3, HER2Retargeting of T Phase I/II Advanced solid (Neovii Biotech, Triomabcells to tumor tumors Fresenius Biotech) Blinatumomab BiTE CD3, CD19Retargeting of T Approved in Precursor B-cell (Blincyto ®, AMG cells totumor USA ALL 103, MT 103, Phase II and ALL MEDI 538, III DLBCL Amgen)Phase II NHL Phase I REGN1979 BsAb CD3, CD20 (Regeneron) Solitomab (AMGBiTE CD3, Retargeting of T Phase I Solid tumors 110, MT110, EpCAM cellsto tumor Amgen) MEDI 565 (AMG BiTE CD3, CEA Retargeting of T Phase IGastrointestinal 211, MedImmune, cells to tumor adenocancinoma Amgen)RO6958688 BsAb CD3, CEA (Roche) BAY2010112 BiTE CD3, PSMA Retargeting ofT Phase I Prostate cancer (AMG 212, Bayer; cells to tumor Amgen) MGD006DART CD3, CD123 Retargeting of T Phase I AML (Macrogenics) cells totumor MGD007 DART CD3, gpA33 Retargeting of T Phase I Colorectal cancer(Macrogenics) cells to tumor MGD011 DART CD19, CD3 (Macrogenics)SCORPION BsAb CD3, CD19 Retargeting of T (Emergent cells to tumorBiosolutions, Trubion) AFM11 (Affimed TandAb CD3, CD19 Retargeting of TPhase I NHL and ALL Therapeutics) cells to tumor AFM12 (Affimed TandAbCD19, CD16 Retargeting of NK Therapeutics) cells to tumor cells AFM13(Affimed TandAb CD30, Retargeting of NK Phase II Hodgkin's Therapeutics)CD16A cells to tumor Lymphoma cells GD2 (Barbara Ann T cells CD3, GD2Retargeting of T Phase I/II Neuroblastoma Karmanos Cancer preloadedcells to tumor and Institute) with BsAb osteosarcoma pGD2 (Barbara Tcells CD3, Her2 Retargeting of T Phase II Metastatic breast Ann Karmanospreloaded cells to tumor cancer Cancer Institute) with BsAb EGFRBi-armedT cells CD3, EGFR Autologous Phase I Lung and other autologous preloadedactivated T cells solid tumors activated T cells with BsAb toEGFR-positive (Roger Williams tumor Medical Center) Anti-EGFR-armed Tcells CD3, EGFR Autologous Phase I Colon and activated T-cells preloadedactivated T cells pancreatic (Barbara Ann with BsAb to EGFR-positivecancers Karmanos Cancer tumor Institute) rM28 (University Tandem CD28,Retargeting of T Phase II Metastatic Hospital Tübingen) scFv MAPG cellsto tumor melanoma IMCgp100 ImmTAC CD3, peptide Retargeting of T PhaseI/II Metastatic (Immunocore) MHC cells to tumor melanoma DT2219ARL 2scFv CD19, CD22 Targeting of Phase I B cell leukemia (NCI, University oflinked to protein toxin to or lymphoma Minnesota) diphtheria tumor toxinXmAb5871 BsAb CD19, (Xencor) CD32b NI-1701 BsAb CD47, CD19 (NovImmune)MM-111 BsAb ErbB2, (Merrimack) ErbB3 MM-141 BsAb IGF-1R, (Merrimack)ErbB3 NA (Merus) BsAb HER2, HER3 NA (Merus) BsAb CD3, CLEC12A NA (Merus)BsAb EGFR, HER3 NA (Merus) BsAb PD1, undisclosed NA (Merus) BsAb CD3,undisclosed Duligotuzumab DAF EGFR, Blockade of 2 Phase I and II Headand neck (MEHD7945A, HER3 receptors, ADCC Phase II cancer Genentech,Roche) Colorectal cancer LY3164530 (Eli Not EGFR, MET Blockade of 2Phase I Advanced or Lily) disclosed receptors metastatic cancer MM-111HSA body HER2, Blockade of 2 Phase II Gastric and (Merrimack HER3receptors Phase I esophageal Pharmaceuticals) cancers Breast cancerMM-141, IgG-scFv IGF-1R, Blockade of 2 Phase I Advanced solid (MerrimackHER3 receptors tumors Pharmaceuticals) RG7221 CrossMab Ang2, VEGFABlockade of 2 Phase I Solid tumors (RO5520985, proangiogenics Roche)RG7716 (Roche) CrossMab Ang2, VEGFA Blockade of 2 Phase I Wet AMDproangiogenics OMP-305B83 BsAb DLL4/VEGF (OncoMed) TF2 Dock and CEA, HSGPretargeting Phase II Colorectal, (Immunomedics) lock tumor for PET orbreast and lung radioimaging cancers ABT-981 DVD-Ig IL-1α, IL-1βBlockade of 2 Phase II Osteoarthritis (AbbVie) proinflammatory cytokinesABT-122 DVD-Ig TNF, IL-17A Blockade of 2 Phase II Rheumatoid (AbbVie)proinflammatory arthritis cytokines COVA322 IgG-fynomer TNF, IL17ABlockade of 2 Phase I/II Plaque psoriasis proinflammatory cytokinesSAR156597 Tetravalent IL-13, IL-4 Blockade of 2 Phase I Idiopathic(Sanofi) bispecific proinflammatory pulmonary tandem IgG cytokinesfibrosis GSK2434735 Dual- IL-13, IL-4 Blockade of 2 Phase I (Healthy(GSK) targeting proinflammatory volunteers) domain cytokinesOzoralizumab Nanobody TNF, has Blockade of Phase II Rheumatoid (ATN103,Ablynx) proinflammatory arthritis cytokine, binds to HSA to increasehalf-life ALX-0761 (Merck Nanobody IL-17A/F, Blockade of 2 Phase I(Healthy Serono, Ablynx) has proinflammatory volunteers) cytokines,binds to HSA to increase half-life ALX-0061 Nanobody IL-6R, has Blockadeof Phase I/II Rheumatoid (AbbVie, Ablynx; proinflammatory arthritiscytokine, binds to HSA to increase half-life ALX-0141 Nanobody RANKL,Blockade of bone Phase I Postmenopausal (Ablynx, has resorption, bindsbone loss Eddingpharm) to HSA to increase half-life RG6013/ACE910 ART-IgFactor IXa, Plasma Phase II Hemophilia (Chugai, Roche) factor Xcoagulation

While this invention has been disclosed with reference to specificaspects, it is apparent that other aspects and variations of thisinvention may be devised by others skilled in the art without departingfrom the true spirit and scope of the invention. The appended claims areintended to be construed to include all such aspects and equivalentvariations.

What is claimed is:
 1. A rupture disk comprising: a body including aninternal wall extending from a first end of the body to a second end ofthe body, the internal wall defining an opening extending through thebody, the body further including an internal flange formed on theinternal wall, the internal flange having a stepped surface; a rupturemembrane positioned within the body, the rupture membrane including arupture membrane surface secured to the stepped surface to form ahermetic seal, the stepped surface and the rupture membrane beingpositioned within the body such that the rupture membrane does notextend beyond the first end of the body; and a metal-to-metal bondbetween a first metal of the internal flange and a second metal of therupture membrane.
 2. The rupture disk of claim 1, wherein the body isconfigured for removable attachment to a component.
 3. The rupture diskof claim 1, wherein the body includes a first metal and the rupturemembrane is fabricated from a second metal, wherein the first metal isselected from 316L stainless steel, Inconel, indium, aluminum, copper,lead, zinc, nickel, silver, palladium, cadmium, titanium, tungsten, tin,gold, and lead, and wherein the second metal is selected from 316Lstainless steel, Inconel, indium, aluminum, copper, lead, zinc, nickel,silver, palladium, cadmium, titanium, tungsten, tin, gold, and lead. 4.The rupture disk of claim 1, wherein the metal-to-metal bond joins thestepped surface of the internal flange and the rupture membrane surfaceof the rupture membrane.
 5. The rupture disk of claim 1, wherein a coldweld joins the stepped surface of the internal flange and the rupturemembrane surface of the rupture membrane.
 6. The rupture disk of claim1, wherein the rupture disk is configured to form a barrier that sealsan aperture of a component to which the rupture disk is attached, andwherein the rupture membrane, prior to rupture, prevents flow of a fluidout of the aperture of the component.
 7. The rupture disk of claim 6,wherein the aperture is greater than 10 mm.
 8. The rupture disk of claim6, wherein the component is suitable for use in the production of abiological product.
 9. The rupture disk of claim 1, wherein the body isconfigured for attachment to a component by a connector.
 10. The rupturedisk of claim 1, wherein the rupture disk is configured to be disposedbetween a first and second component, and wherein the rupture membrane,prior to rupture, forms a barrier between the first component and thesecond component.
 11. The rupture disk of claim 1, wherein the body isconfigured as a spool piece.
 12. The rupture disk of claim 1, whereinthe body is configured so as to be removably attached by a clamp to acomponent.
 13. The rupture disk of claim 1, further comprising a gasketconfigured to be disposed between the first end of the body and acomponent and to provide a fluid-proof seal there between.
 14. Therupture disk of claim 1, wherein the rupture membrane is positioned suchthat it is entirely within the body.
 15. The rupture disk of claim 1,wherein the rupture membrane is positioned such that it is entirelywithin the body, but close enough to the first end of the housing tominimize a volume defined by a clean surface of the rupture membrane anda portion of the internal wall between the clean surface of the rupturemembrane and the first end of the body.
 16. The rupture disk of claim 1,wherein the rupture membrane is positioned such that it is entirelywithin the body such that a volume of a space defined by a face of therupture membrane and a portion of the internal wall between the face ofthe rupture membrane and the first end of the body is no more than 10cm³.
 17. The rupture disk of claim 1, wherein the internal flange andthe internal wall are integrally formed.
 18. The rupture disk of claim1, the rupture disk consisting of the rupture membrane and the bodyformed as a spool piece.
 19. The rupture disk of claim 1, the rupturedisk consisting essentially of the rupture membrane and the body formedas a spool piece.
 20. A process system comprising a rupture disk ofclaim 1, wherein the process system includes a bioreactor.
 21. Theprocess system comprising a rupture disk of claim 1, wherein the processsystem is a bioprocess system.
 22. A method of maintaining a processsystem comprising: detaching a rupture disk of claim 1, from acomponent; and reattaching the rupture disk to the component, therebymaintaining the process system.
 23. The method of claim 22, wherein theprocess system is a bioprocess system.